(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2006/0172944 A1
`Wiegand et al.
`(43) Pub. Date:
`Aug. 3, 2006
`
`US 2006O172944A1
`
`(54) METHOD OF TREATING EYE INJURY
`WITH LOCAL ADMINISTRATION OF A
`VEGF INHIBITOR
`(76) Inventors: Stanley J. Wiegand,
`Croton-on-Hudson, NY (US); Jingtai
`Cao, Chappaqua, NY (US)
`Correspondence Address:
`REGENERON PHARMACEUTICALS, INC
`777 OLD SAW MILL RIVER ROAD
`TARRYTOWN, NY 10591 (US)
`(21) Appl. No.:
`11/346,009
`
`(22) Filed:
`
`Feb. 2, 2006
`Related U.S. Application Data
`(60) Provisional application No. 60/649.232, filed on Feb.
`2, 2005.
`
`Publication Classification
`
`(51) Int. Cl.
`(2006.01)
`A6II 38/17
`(52) U.S. Cl. ................................................................ S14/12
`
`(57)
`
`ABSTRACT
`
`Methods of reducing or treating angiogenesis and/or inflam
`mation associated with eye injury in a subject in need
`thereof, comprising administering an agent capable of
`blocking or inhibiting vascular endothelial growth factor
`(VEGF) are provided. The methods are useful for inhibiting
`or ameliorating eye injury, particularly acute or Subacute
`corneal injury and feature local administration (for example,
`Subconjunctival injection or eye drops).
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 1
`
`Joining Petitioner: Apotex
`
`

`

`Patent Application Publication Aug. 3, 2006 Sheet 1 of 7
`
`US 2006/0172944 A1
`
`
`
`
`
`s
`
`ce
`er
`
`ces
`n
`
`s
`
`ed
`
`n
`
`(%) eelW Jeno SeA
`
`
`
`C
`r
`
`es
`r
`
`C
`n
`
`ed
`
`g
`
`CC
`
`(%) eelW Jenso SeW
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 2
`
`Joining Petitioner: Apotex
`
`

`

`Patent Application Publication Aug. 3, 2006 Sheet 2 of 7
`
`US 2006/0172944 A1
`
`
`
`f
`
`C
`r
`
`C
`ty
`
`C
`n
`
`O
`d
`
`C
`
`(%) eeuw enoSeA
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 3
`
`Joining Petitioner: Apotex
`
`

`

`
`
`(un) fuel esse A
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 4
`
`Joining Petitioner: Apotex
`
`

`

`Patent Application Publication Aug. 3, 2006 Sheet 4 of 7
`
`US 2006/0172944 A1
`
`S.
`O
`D
`Qo
`O
`u
`H
`L
`CD
`D
`
`O
`
`E -
`as O
`(f)
`?o
`5
`
`5
`Z
`
`vs.
`d
`in
`
`
`
`O
`O
`N
`
`O
`lf)
`V
`
`O
`O
`V
`
`O
`L)
`
`(un) up fuel essew
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 5
`
`Joining Petitioner: Apotex
`
`

`

`Patent Application Publication Aug. 3, 2006 Sheet 5 of 7
`
`US 2006/0172944 A1
`
`
`
`:
`
`2 .
`
`s
`
`O
`O
`O
`O
`f
`(f)
`N
`(Y)
`(un) sseuxou, eeuoso
`
`O
`O
`GN
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 6
`
`Joining Petitioner: Apotex
`
`

`

`Patent Application Publication Aug. 3, 2006 Sheet 6 of 7
`
`US 2006/0172944 A1
`
`6nG
`
`6nOOI
`
`6nOOI
`
`(OqnS) deu ? JOBA
`
`9 -61-I
`
`
`
`(OS) del L JEDEA
`
`0/607
`
`0/60
`
`0/60 [
`
`9/008
`
`9/009
`
`9/0017
`
`o/ Uoon pe
`
`
`
`9/008
`
`9/60/
`
`9/009
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 7
`
`Joining Petitioner: Apotex
`
`

`

`Patent Application Publication Aug. 3, 2006 Sheet 7 of 7
`
`US 2006/0172944 A1
`
`
`
`i
`
`f
`
`O
`O
`
`O O O O O
`O O O O
`V
`?y
`N
`Y
`
`ufuel esseA
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 8
`
`Joining Petitioner: Apotex
`
`

`

`US 2006/0172944 A1
`
`Aug. 3, 2006
`
`METHOD OF TREATING EYE NURY WITH
`LOCAL ADMINISTRATION OF A VEGF
`INHIBITOR
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`0001) This application claims the benefit under 35 USC S
`119(e) of U.S. Provisional 60/649,232 filed 2 February 2005,
`which application is herein specifically incorporated by
`reference in its entirety.
`
`BACKGROUND
`0002) 1. Field of the Invention
`0003. The field of the invention is related to local admin
`istration of VEGF antagonists to treat eye-related diseases,
`disorders and injuries.
`0004 2. Description of Related Art
`0005. It has previously been reported that topical appli
`cation of an anti-VEGF neutralizing antibody suppresses
`acute allograft rejection in a rat corneal transplant model
`(Yatoh et al. (1998) Transplantation 66(11): 1519-24).
`
`BRIEF SUMMARY OF THE INVENTION
`0006 The invention is based in part on the finding that
`local administration of an agent capable of blocking, inhib
`iting, or reducing the activity of vascular endothelial growth
`factor (VEGF) is useful in treating of angiogenesis and
`inflammation associated with eye injuries or infection.
`0007. In a first aspect, the invention features a method of
`treating an eye injury, comprising locally administering an
`effective amount of an agent capable of blocking or inhib
`iting vascular endothelial growth factor (VEGF)-mediated
`activity to a subject in need thereof. Such that the eye injury
`is ameliorated or improved. Preferably, the eye injury is a
`corneal injury or conjunctival injury and the method of
`treatment reduces angiogenesis and inflammation associated
`with the eye injury.
`0008. In specific embodiments, the agent capable of
`blocking, inhibiting, or ameliorating VEGF-mediated activ
`ity is a VEGF antagonist comprising a fusion polypeptide
`selected from the group consisting of acetylated Flt-1 (1-3)-
`Fc, Flt-1 (1-3A)-Fc, Fit-1 (1-3A)-Fc, Flt-1 (2-3A)-Fc, Flt
`1(2-3)-Fc, Flt-1 D2-VEGFR3D3-FcAC1 (a), Flt-1 D2-Flk-1
`D3-FcAC1 (a), and VEGFR1R2-FcAC1 (a). In a specific and
`preferred embodiment, the VEGF trap is VEGFR1
`R2-FcAC1 (a) (also termed VEGF trap) comprising the
`nucleotide sequence set forth in SEQ ID NO: 1 and the
`amino acid sequence set forth in SEQ ID NO: 2. The
`invention comprises the use of a VEGF trap that is at least
`90%. 95%, 98%, or at least 99% homologous with the
`nucleotide sequence set forth in SEQ ID NO: 1 and/or the
`amino acid sequence set forth in SEQ ID NO:2.
`0009. The method of the invention is useful to treat acute
`and Sub-acute corneal injury or conjunctival injury. Acute
`corneal injury may be treated within 24 hours of occurrence,
`and includes corneal injury or conjunctival injury caused by
`a penetrating object, a foreign body, or a chemical or burn
`injury. A sub-acute injury may be treated up to two weeks
`post-injury and may include the above listed injuries as well
`as infectious etiologies.
`
`0010. In various embodiments, the eye injury is caused
`by trauma, e.g., Surgical injuries, chemical burn, corneal
`transplant, infectious or inflammatory diseases.
`0011 Length of treatment will vary according to the
`injury, but treatment duration may be short, e.g., up to one
`month, and may include a 3-6 month observation period,
`during which retreatment may be provided.
`0012 Administration may also include a second agent,
`Such as an immunosuppressive agent, for example, one or
`more of a corticosteroid, dexamethasone, or cyclosporin A.
`0013 Local administration includes, for example, admin
`istration of the VEGF antagonist in eye drops applied to the
`eye, or Subconjunctival injection to the eye.
`0014. In a second aspect, the invention features a method
`of healing an eye injury, comprising locally administering an
`effective amount of an agent capable of blocking or inhib
`iting vascular endothelial growth factor (VEGF)-mediated
`activity to a subject in need thereof. Such that the eye injury
`heals.
`0015. In a third aspect, the invention features a method of
`reducing or ameliorating angiogenesis associated with an
`eye injury, comprising locally administering an effective
`amount of an agent capable of blocking or inhibiting vas
`cular endothelial growth factor (VEGF)-mediated activity to
`a Subject in need thereof. Such that the angiogenesis asso
`ciated with the eye injury is reduced or ameliorated.
`0016.
`In a fourth aspect, the invention features a method
`of reducing or ameliorating inflammation associated with an
`eye injury, comprising locally administering an effective
`amount of an agent capable of blocking or inhibiting vas
`cular endothelial growth factor (VEGF)-mediated activity to
`a Subject in need thereof. Such that the inflammation asso
`ciated with the eye injury is reduced or ameliorated.
`0017. In a fifth aspect, the invention features an oph
`thalmic composition comprising a VEGF antagonist, for
`example the VEGF trap VEGFR1R2-FcAC1 (a), in a phar
`maceutically acceptable carrier. Such pharmaceutical com
`positions may be liquid, gel, ointment, salve, slow release
`formulations or other formulations suitable for ophthalmic
`administration. In various embodiments, the pharmaceutical
`composition is for local administration comprising a VEGF
`trap, buffer, Stabilizer, isotonizer, and a pharmaceutical
`carrier. In a preferred embodiment, the pharmaceutical com
`position is administered in the form of eye drops. In specific
`embodiments, the pharmaceutically acceptable carrier com
`prises as least one ophthalmically acceptable excipient,
`wherein the ophthalmically acceptable excipient can reduce
`a rate of removal of the VEGF antagonist from the eye by
`lacrimation. In various preferred embodiments, the pharma
`ceutical composition has an effective residence time in the
`eye of about 2 to about 24 hours.
`0018. In other embodiments, the pharmaceutical compo
`sition is for Subconjuctival administration Such as Subcon
`juctival injection and Subconjuctival implantation.
`0019. In a sixth aspect, the invention features a method of
`administering a VEGF antagonist for treatment of angio
`genesis and/or inflammation associated with eye injury or
`infection, comprising local administration by eye drops
`comprising a VEGF trap, or Subconjunctival administration
`by injection or implantation.
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 9
`
`Joining Petitioner: Apotex
`
`

`

`US 2006/0172944 A1
`
`Aug. 3, 2006
`
`0020. Other objects and advantages will become apparent
`from a review of the ensuing detailed description.
`
`BRIEF DESCRIPTION OF THE FIGURES
`FIG. 1. Percent of vascularized corneal area in
`0021
`sutured mice subconjunctivally (SubC) treated with vehicle
`only or VEGF trap, at dosing regimens of (A) three 40 ug or
`(B) three 10 ug doses.
`0022 FIG. 2. Percent of neovascularized cornea at day 9
`in sutured rats treated SubC with vehicle only or treated with
`VEGF trap, with a dosing regimen of 10 ug on the day of
`Suturing.
`0023 FIG. 3. Blood vessel length in sutured rats treated
`at day 0, 3 and 6 with 25, 50, or 100 ug VEGF trap injections
`administered subcutaneously (SC) or subconjunctivally
`(SubC). (Control =right non-sutured eye).
`0024 FIG. 4. Quantification of blood vessel length in
`Sutured rats receiving Subconjunctival normal Saline, 5 Jug,
`25 ug, or 100 lug. VEGF trap on day 0, 3 and 6. (Control
`=right non-Sutured eye).
`0.025
`FIG. 5. Quantification of corneal edema as evi
`denced by corneal thickness in Sutured rats receiving 25 or
`100 ug VEGF trap SC or normal saline, 5ug. 25 ug, or 100
`ug VEGF trap SubC. (Control =right non-sutured eye)
`(NS=sutured eye, normal saline administered SubC).
`0026 FIG. 6. Percent reduction of edema. Effect of
`VEGF trap on inflammation as determined by measurement
`of corneal thickness. All animals were sutured (control
`=Sutured +systemic injection (SC) of normal saline).
`0027 FIG. 7. Blood vessel length in suture-injury. Con
`trol =no Suture injury. Suture control =Suture--no treatment.
`Vehicle=suture-injury--vehicle provided as eye drops. VEGF
`trap =Suture-injury--1 drop three times per day (412 jug
`VEGF trap protein/drop).
`
`DETAILED DESCRIPTION
`0028 Before the present methods are described, it is to be
`understood that this invention is not limited to particular
`methods, and experimental conditions described, as Such
`methods and conditions may vary. It is also to be understood
`that the terminology used herein is for the purpose of
`describing particular embodiments only, and is not intended
`to be limiting, since the scope of the present invention will
`be limited only by the appended claims.
`0029. As used in this specification and the appended
`claims, the singular forms “a”, “an', and “the include plural
`references unless the context clearly dictates otherwise.
`Thus for example, a reference to “a method’ includes one or
`more methods, and/or steps of the type described herein
`and/or which will become apparent to those persons skilled
`in the art upon reading this disclosure and so forth.
`0030 Unless defined otherwise, all technical and scien
`tific terms used herein include the same meaning as com
`monly understood by one of ordinary skill in the art to which
`this invention belongs. Although any methods and materials
`similar or equivalent to those described herein can be used
`in the practice or testing of the present invention, preferred
`methods and materials are now described. All publications
`mentioned herein are incorporated herein by reference in
`their entirety.
`
`0.031) General Description
`0032 Experiments were undertaken to evaluate corneal
`neovascularization after Surgical Suture placement in the
`cornea and to test whether corneal neovascularization fol
`lowing Suture injury can be Suppressed by local administra
`tion of an agent capable of blocking, inhibiting, or amelio
`rating VEGF-mediated activity. As described in the
`experimental section below, corneas of male C57BL/6 mice
`or Sprague-Dawley rats were suture-injured. A molecular
`trap designed to inhibit VEGF-A activity was administered
`locally and tested for its ability to suppress corneal vascu
`larization. The results revealed that sutured cornea receiving
`subconjunctival administration of VEGF trap exhibited little
`or no neovascularization; corneal vascular area and vessel
`length following Suture injury being comparable to that of a
`normal untreated (nonsutured) cornea. Treatment with
`VEGF trap eye drops following suture injury also effectively
`reduced neovascularization in Suture-injured cornea.
`0033. In addition to quantification of neovascularization
`as measured by an increase in either blood vessel length or
`blood vessel area, Suture-injury produced a marked influx of
`leucocytes into the injury site. When VEGF trap was admin
`istered locally either by subconjunctival injection (SubC) or
`by eye drop, a dramatic reduction in leucocyte infiltration
`was observed (data not shown).
`0034. In addition to the measurements reported below,
`serum levels of VEGF trap were determined in animals
`treated by subconjunctival injection or eye drops of VEGF
`trap. As evidenced by ELISA measurement for free VEGF
`trap in serum, there is little or no systemic exposure when
`VEGF trap is delivered at the effective doses by either of
`these local (SubC or eye drops) routes.
`0035) Definitions
`0036) The phrase “therapeutically effective dose”
`includes a dose that produces the desired effect for which it
`is administered. The exact dose will depend on the purpose
`of the treatment, and will be ascertainable by one skilled in
`the art using known techniques (see, for example, Lloyd
`(1999) The Art, Science and Technology of Pharmaceutical
`Compounding).
`0037. The term “blocker”, “inhibitor', or “antagonist”
`are used interchangeably to mean a Substance that retards or
`prevents a chemical or physiological reaction or response.
`Common blockers or inhibitors comprise, but are not limited
`to, antisense molecules, antibodies, antagonists and their
`derivatives. More specifically, an example of a VEGF
`blocker or inhibitor includes a VEGF receptor-based antago
`nist comprising, for example, an anti-VEGF antibody, or a
`VEGF trap such as VEGFR1R2-FcAC1(a) (SEQ ID NOS:1-
`2).
`0038. The phrase “ophthalmically acceptable” with
`respect to a formulation, composition or ingredient herein
`means having no persistent effect that is substantially det
`rimental to the treated eye or the functioning thereof, or on
`the general health of the subject being treated. It will be
`recognized that transient effects such as minor irritation or a
`"stinging sensation are common with topical ophthalmic
`administration of drugs and the existence of Such transient
`effects is not inconsistent with the formulation, composition
`or ingredient in question being “ophthalmically acceptable'
`as herein defined. However, preferred formulations, compo
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 10
`
`Joining Petitioner: Apotex
`
`

`

`US 2006/0172944 A1
`
`Aug. 3, 2006
`
`sitions and ingredients are those that cause no substantial
`detrimental effect, even of a transient nature.
`0039) VEGF Antagonists
`0040. In various embodiments, the VEGF trap is selected
`from the group consisting of acetylated Fit-1 (1-3)-Fc, Fit
`1(1-3A)-Fc, Flt-1 (1-3AB)-Fc, Flt-1 (2-3AB)-Fc, Flt-1 (2-3)-
`Fc, Flt-1D2-VEGFR3D3-FcAC1 (a), Flt-1D2-Flk-1D3
`FcAC1 (a), and VEGFR1R2-FcAC1 (a). For a more detailed
`description of these and other VEGF-receptor-based antago
`nists, including pegylated receptor-based blockers, see PCT
`WO/00/75319, the contents of which are incorporated in
`their entirety herein by reference.
`0041. In addition to the VEGF receptor-based antagonists
`disclosed in PCT WO/00/75319, which publication is herein
`specifically incorporated by reference in its entirety, variants
`and derivatives of such VEGF receptor-based blockers are
`also contemplated by the invention. The sequence of the
`variants or derivatives may differ by a change that can be
`one or more additions, insertions, deletions and/or substitu
`tions of one or more nucleotides of the sequence set forth in
`SEQID NO: 1 Changes to a nucleotide sequence may result
`in an amino acid change at the protein level, or not, as
`determined by the genetic code. Thus, nucleic acid accord
`ing to the present invention may include a sequence different
`from the sequence shown in SEQ ID NO:1, yet encode a
`polypeptide with the same amino acid sequence as SEQID
`NO: 2. On the other hand, the encoded polypeptide may
`comprise an amino acid sequence which differs by one or
`more amino acid residues from the amino acid sequence
`shown in SEQID NO:2. A nucleic acid encoding a polypep
`tide which is an amino acid sequence variant or derivative of
`the sequence shown in SEQID NO:2 is further provided by
`the present invention. A nucleic acid encoding Such a
`polypeptide may show at the nucleotide sequence and/or
`encoded amino acid level greater than about 90%. 95%,
`98%, or 99% homology with the coding sequence shown in
`SEQ ID NO:1 and/or the amino acid sequence shown in
`SEQID NO:2. Amino acid “homology’, may be understood
`to be similarity (according to the established principles of
`amino acid similarity, e.g. as determined using the algorithm
`GAP (Genetics Computer Group, Madison, Wis.)) or iden
`tity. GAP uses the Needleman and Wunsch algorithm to
`align two complete sequences that maximizes the number of
`matches and minimizes the number of gaps. Generally, the
`default parameters are used, with a gap creation penalty=12
`and gap extension penalty=4.
`0.042
`Individual components of the VEGF-specific
`fusion proteins of the invention may be constructed by
`molecular biological methods known to the art with the
`guidance provided by the instant specification. These com
`ponents are selected from a first cellular receptor protein,
`such as, for example, VEGFR1; a second cellular receptor
`protein, such as, for example, VEGFR2; and a multimerizing
`component, such as, for example, an Fc.
`0.043
`Specific embodiments of the VEGF-specific fusion
`proteins useful in the methods of the invention comprise a
`multimerizing component which allows the fusion proteins
`to associate, e.g., as multimers, preferably dimers. Prefer
`ably, the multimerizing component comprises an immuno
`globulin-derived domain. Suitable multimerizing compo
`nents are sequences encoding an immunoglobulin heavy
`chain hinge region (Takahashi et al. 1982 Cell 29:671-679);
`immunoglobulin gene sequences, and portions thereof.
`
`0044) The nucleic acid constructs encoding the fusion
`proteins useful in the methods of the invention can be
`inserted into an expression vector by methods known to the
`art, wherein the nucleic acid molecule can be operatively
`linked to an expression control sequence. Host-vector sys
`tems for the production of proteins comprising an expression
`vector introduced into a host cell suitable for expression of
`the protein are known in the art. The suitable host cell may
`be a bacterial cell Such as E. coli, a yeast cell. Such as, for
`example, Pichia pastoris, an insect cell. Such as, for
`example, Spodoptera frugiperda, or a mammalian cell. Such
`as, for example, a COS, CHO, 293, BHK or NSO cell.
`0045 Methods of Administration
`0046) The invention comprises methods of treatment
`comprising administering to a subject an effective amount of
`an agent of the invention. In a preferred aspect, the agent is
`substantially purified (e.g., substantially free from sub
`stances that limit its effect or produce undesired side
`effects). The Subject is preferably an animal, e.g., such as
`cows, pigs, horses, chickens, cats, dogs, etc., and is prefer
`ably a mammal, and most preferably human.
`0047 Preferably, the pharmaceutical compositions of the
`invention are administered to the area in need of treatment
`by topical administration. Topical drug delivery is the most
`common treatment for diseases or disorders of the anterior
`segment of the eye, including, for example, corneal diseases,
`uveitis, and glaucoma. Topical delivery can be a safer and
`more convenient delivery method for patients, and can
`reduce the risk of many side effects observed in systemic
`treatment regimens. Topical administration of an angiogen
`esis inhibitor to the eye or cornea can be an effective
`treatment for treating neovascularization and/or inflamma
`tion. A preferred method of administering the pharmaceuti
`cal compositions of the invention to the eye is by eye drops
`comprising a VEGF trap.
`0048. In various preferred embodiments, the pharmaceu
`tical compositions of the invention are administered to the
`area in need of treatment by Subconjunctival administration.
`One preferred method of subconjunctival administration to
`the eye is by injectable formulations comprising a VEGF
`trap. Another preferred method of subconjunctival admin
`istration is by implantations comprising slow releasing
`VEGF trap.
`0049 Pharmaceutical Compositions
`0050 Pharmaceutical compositions useful in the practice
`of the method of the invention include a therapeutically
`effective amount of an active agent with a pharmaceutically
`acceptable carrier. The term “pharmaceutically acceptable'
`means approved by a regulatory agency of the Federal or a
`state government or listed in the U.S. Pharmacopeia or other
`generally recognized pharmacopeia for use in animals, and
`more particularly, in humans. The term “carrier refers to a
`diluent, adjuvant, excipient, or vehicle with which the thera
`peutic is administered. Examples of Suitable pharmaceutical
`carriers are described in “Remington’s Pharmaceutical Sci
`ences” by E.W. Martin. In a preferred embodiment, the
`composition is formulated in accordance with routine pro
`cedures as a pharmaceutical composition adapted for topical
`administration to human beings. Such pharmaceutical com
`positions may be liquid, gel, ointment, salve, slow release
`formulations or other formulations suitable for ophthalmic
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 11
`
`Joining Petitioner: Apotex
`
`

`

`US 2006/0172944 A1
`
`Aug. 3, 2006
`
`administration. The composition comprises an effective
`amount of VEGF antagonist and, optionally, at least one
`ophthalmically acceptable excipient, wherein the excipient
`is able to reduce a rate of removal of the composition from
`the eye by lacrimation, such that the composition has an
`effective residence time in the eye of about 2 hours to about
`24 hours.
`0051. In various embodiments, compositions of the
`invention can comprise a liquid comprising an active agent
`in solution, in Suspension, or both. The term 'suspension”
`herein includes a liquid composition wherein a first portion
`of the active agent is present in Solution and a second portion
`of the active agent is present in particulate form, in Suspen
`sion in a liquid matrix. As used herein, liquid compositions
`include gels.
`0.052
`Preferably the liquid composition is aqueous.
`Alternatively, the composition can take form of an ointment.
`In a preferred embodiment, the composition is an in situ
`gellable aqueous composition, more preferably an in situ
`gellable aqueous Solution. Such a composition can comprise
`a gelling agent in a concentration effective to promote
`gelling upon contact with the eye or lacrimal fluid in the
`exterior of the eye. Suitable gelling agents non-restrictively
`include thermosetting polymers such as tetra-substituted
`ethylene diamine block copolymers of ethylene oxide and
`propylene oxide (e.g., poloxamine 1307); polycarbophil;
`and polysaccharides such as gellan, carrageenan (e.g.,
`kappa-carrageenan and iota-carrageenan), chitosan and algi
`nate gums. The phrase “in situ gellable' includes not only
`liquids of low viscosity that can form gels upon contact with
`the eye or with lacrimal fluid in the exterior of the eye, but
`also more viscous liquids such as semi-fluid and thixotropic
`gels that exhibit Substantially increased viscosity or gel
`stiffness upon administration to the eye or area Surrounding
`the eye.
`0053 Aqueous compositions of the invention have oph
`thalmically compatible pH and osmolality. Preferably these
`compositions incorporate means to inhibit microbial growth,
`for example through preparation and packaging under sterile
`conditions and/or through inclusion of an antimicrobially
`effective amount of an ophthalmically acceptable preserva
`tive. Suitable preservatives non-restrictively include mer
`cury-containing Substances such as phenylmercuric salts
`(e.g., phenylmercuric acetate, borate and nitrate) and thime
`rosal; Stabilized chlorine dioxide; quaternary ammonium
`compounds such as benzalkonium chloride, cetyltrimethy
`lammonium bromide and cetylpyridinium chloride; imida
`Zolidinyl urea; parabens such as methylparaben, ethylpara
`ben, propylparaben and butylparaben, and salts thereof.
`phenoxyethanol; chlorophenoxyethanol; phenoxypropanol:
`chlorobutanol; chlorocresol; phenylethyl alcohol; disodium
`EDTA; and sorbic acid and salts thereof.
`0054 The composition can comprise an ophthalmic
`depot formulation comprising an active agent for Subcon
`junctival administration. The ophthalmic depot formulation
`comprises microparticles of essentially pure active agent,
`e.g., the VEGF trap of SEQ ID NO:2. The microparticles
`comprising VEGF trap can be embedded in a biocompatible
`pharmaceutically acceptable polymer or a lipid encapsulat
`ing agent. The depot formulations may be adapted to release
`all of substantially all the active material over an extended
`period of time. The polymer or lipid matrix, if present, may
`
`be adapted to degrade sufficiently to be transported from the
`site of administration after release of all or substantially all
`the active agent. The depot formulation can be liquid for
`mulation, comprising a pharmaceutical acceptable polymer
`and a dissolved or dispersed active agent. Upon injection,
`the polymer forms a deot at the injections site, e.g. by
`gelifying or precipitating.
`0055. The composition can comprise a solid article that
`can be inserted in a suitable location in the eye. Such as
`between the eye and eyelid or in the conjunctival sac, where
`the article releases the active agent. Release from Such an
`article is preferably to the cornea, either via lacrimal fluid
`that bathes the surface of the cornea, or directly to the cornea
`itself, with which the solid article is generally in intimate
`contact. Solid articles suitable for implantation in the eye in
`Such fashion generally comprise polymers and can be bio
`erodible or non-bioerodible. Bioerodible polymers that can
`be used in preparation of ocular implants carrying a VEGF
`trap in accordance with the present invention include with
`out restriction aliphatic polyesters such as polymers and
`copolymers of poly(glycolide), poly(lactide), poly(e-capro
`lactone), poly(hydroxybutyrate) and poly(hydroxyvalerate),
`polyamino acids, polyorthoesters, polyanhydrides, aliphatic
`polycarbonates and polyether lactones. Illustrative of Suit
`able non-bioerodible polymers are silicone elastomers.
`0056. The active agents of the invention can be formu
`lated as neutral or salt forms. Pharmaceutically acceptable
`salts include those formed with free amino groups such as
`those derived from hydrochloric, phosphoric, acetic, oxalic,
`tartaric acids, etc., and those formed with free carboxyl
`groups such as those derived from Sodium, potassium,
`ammonium, calcium, ferric hydroxides, isopropylamine, tri
`ethylamine, 2-ethylamino ethanol, histidine, procaine, etc.
`0057 Combination Therapies
`0058. In various embodiments, the VEGF antagonists of
`the present invention may be administered in combination
`with one or more additional compounds or therapies or
`medical procedures. For example, Suitable therapeutic
`agents for use in combination, either alternating or simul
`taneously, with the VEGF-binding fusion proteins of the
`invention, including topically administered immunosuppres
`sive agents such as corticosteroids, dexamethasone,
`cyclosporin A, FK506, or anti-metabolic agents, (see Barker,
`NH, et al., (2000) Clin Exp Opthal 28:357-360). Other
`Suitable therapeutic agents for use in combination, either
`alternating or simultaneously, with the VEGFantagonists of
`the invention may include agents that can block the biologi
`cal activity of other VEGF family members such as VEGF-C
`and VEGF-D.
`0059) Kits
`0060. The invention also provides an article of manufac
`ture comprising packaging material and a pharmaceutical
`agent contained within the packaging material, wherein the
`pharmaceutical agent comprises at least one VEGF-specific
`fusion protein of the invention and wherein the packaging
`material comprises a label or package insert that indicates
`that the VEGF-specific fusion protein can be used for
`treating eye injury. The kit can comprise a composition
`comprising a VEGF trap and one or more other components
`Such as, for example, components to be combined prior to
`use either by a health care professional or by the subject. In
`
`Mylan Exhibit 1084
`Mylan v. Regeneron, IPR2021-00881
`Page 12
`
`Joining Petitioner: Apotex
`
`

`

`US 2006/0172944 A1
`
`Aug. 3, 2006
`
`one embodiment, the VEGF trap is combined with one or
`more components that can comprise, for example, a Solution
`included in the kit to reconstitute a VEGF trap in the form
`of an ophthalmical composition Suitable for topical or Sub
`conjunctival administration to a human or animal. Kit com
`ponents can comprise, for example, normal saline solutions
`and/or solutions comprising one or more Suitable pharma
`ceutical carriers, stabilizers, additives, or buffers. Preferably
`the kit comprises instructions for treatment or administration
`regimens and/or instructions for preparing or reconsitituting
`a VEGF trap for use. The instructions can be in writing on
`paper, on computer media of any suitable type, as audiovi
`sual materials including, for example, CD or DVD, or any
`other suitable format.
`0061. Other features of the invention will become appar
`ent in the course of the following descriptions of exemplary
`embodiments which are given for illustration of the inven
`tion and are not intended to be limiting thereof.
`
`EXAMPLES
`0062) The following example is put forth so as to provide
`those of ordinary skill in the art with a complete disclosure
`and description of how to make and use the methods and
`compositions of the invention, and are not intended to limit
`the scope of what the inventors regard as their invention.
`Efforts have been made to ensure accuracy with respect to
`numbers used (e.g., amounts, temperature, etc.) but some
`experimental errors and deviations should be accounted for.
`Unless indicated otherwise, parts are parts by weight,
`molecular weight is average molecular weight, temperature
`is in degrees Centigrade, and pressure is at or near atmo
`spheric.
`
`Example 1
`
`Effect of Eye Drop Administration of VEGF Trap
`on Corneal Neovascularization.
`0063 Topical administration at the cornea of a VEGF
`trap can effectively inhibit or reduce corneal neovascular
`ization and/or inflammation of an injured cornea. Corneal
`injuries, such as those Sustained on repeat penetrating
`keratoplasty and corneal graft procedures, can lead to
`inflammation and/or neovascularization of the cornea, some
`times resulting in corneal graft rejection.
`0064. A study of corneal neovascularization following
`Suture injury in the presence and absence of top